Natural Glycans for Functional Glycomics Project Summary/ Abstract During the first decade of the 21st century, NIH made significant investments in glycomics. The Consortium for Functional Glycomics (supported by NIGMS 2001-2012) led a major international effort using a microarray of 611 defined glycans to identify the protein-glycan interactions associated with normal physiology and disease (http://www.functionalglycomics.org/CFGparadigms/index.php/Welcome_to_the_CFG_Paradigm_Pages). This 611 glycan library transformed studies on protein-glycan interactions and revealed the importance of defining the human glycome, estimated to be in the range of 10,000 determinants. A microarray comprised of all possible glycans would allow us to detect and define all physiologically relevant glycan binding proteins (GBP), but the CFG library is now only 575 glycans making the production of large quantities of pure, defined glycans for functional studies critical for advancing glycomics; e.g, mgs of pure human milk oligosaccharides (HMO), available since the 1950?s supported research that led to our understanding of the benefits of HMO in breasfed infants, resulting in infant formula manufacturers introducing HMO into their products in 2014, based on clinical trials supporting their efficacy. Mammalian glycans,on the other hand, expressed on the surfaces of all cells, are generally available only in minute quantities; making them as available as HMO to the research community would rapidly advance understanding of their functions in animal cells. This is now possible using our proprietary technology, Oxidative Release of Natural Glycans (ORNG), which chemically releases glycans from glycoconjugates of any natural material using household bleach. NatGlycan, LLC (NGC) was established to provide biologically relevant natural glycans reliably, economically, and in large quantities. In Phase I we demonstrated the feasibility of producing high mannose N-glycans in mg quantities for sale to the glycoscience community for the first time. We believe the economy of scale will support reasonable prices. Sigma-Aldrich sells 20g of Man9GlcNAc2 for $688, while NGC is selling 5 mg of Man9GlcNAc2-AEAB for $750. In Phase II of this project we will complete a manufacturing facility to produce 100?s of mg (purity >95%) of all high mannose N- glycans and derivatives that will be useful to the glycobiology community. In addition, we plan to launch highly purified N-glycans representing all of the structures expressed on human and recombinant IgG for use as QC standards in mAb manufacturing; currently an unmet need in the pharmaceutical industry. Since ORNG releases all classes of glycans simultaneously from natural products, we will rapidly expand our product offerings. We will purify the major O-glycans that are in great demand, but not currently available at reasonable cost. We anticipate that by the end of Phase II, we will be offering hundreds of N-glycans and O-glycans (purity >95%) and useful derivatives in multi-miligram amounts. Based on significant sales and orders after our initial product catalog launch, we anticipate being able to support our glycan manufacturing costs and increase profit margins as we increase manufacturing scale in the future.